{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T01:21:57Z","timestamp":1760059317003,"version":"build-2065373602"},"reference-count":23,"publisher":"MDPI AG","issue":"6","license":[{"start":{"date-parts":[[2025,6,4]],"date-time":"2025-06-04T00:00:00Z","timestamp":1748995200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Mauna Kea Technologies"},{"name":"Telix Pharmaceuticals"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pharmaceuticals"],"abstract":"<jats:p>In prostate cancer (PCa) surgery, precise tumor margin identification remains challenging despite advances in surgical techniques. This study evaluates the combination of tumor-specific near-infrared imaging with the PSMA-targeting molecule PSMA-914 and optical endomicroscopy (NIR-pCLE) for single-cell-level tumor identification in a preclinical proof of concept. Methods: NIR-pCLE imaging of varying PSMA-914 concentrations was performed on PSMA-positive LNCaP and PSMA-negative PC-3 cells using Cellvizio\u00ae 100 with pCLE Confocal Miniprobes\u2122. To identify optimal PSMA-914 dosing for in vivo imaging, different doses (0\u201310 nmol) were evaluated using NIR-pCLE, Odyssey CLx imaging, and confocal microscopy in an LNCaP tumor-bearing xenograft model. A proof of concept mimicking a clinical workflow was performed using 5 nmol [68Ga]Ga-PSMA-914 in LNCaP and PC-3 tumor xenografts, including PET\/MRI, in\/ex vivo NIR-pCLE imaging, and microscopic\/macroscopic imaging. Results: NIR-pCLE detected PSMA-specific fluorescence at concentrations above 30 nM in vitro. The optimal dose was identified as 5 nmol PSMA-914 for NIR-pCLE imaging with cellular resolution in LNCaP xenografts. PET\/MRI confirmed high tumor uptake and a favorable distribution profile of PSMA-914. NIR-pCLE imaging enabled real-time, single-cell-level detection of PSMA-positive tissue, visualizing tumor heterogeneity, confirmed by ex vivo microscopy and imaging. Conclusions: This preclinical proof of concept demonstrates the potential of intraoperative PSMA-specific NIR-pCLE imaging to visualize tissue structures in real time at cellular resolution. Clinical implementation could provide surgeons with valuable additional information, potentially advancing PCa patient care through improved surgical precision.<\/jats:p>","DOI":"10.3390\/ph18060841","type":"journal-article","created":{"date-parts":[[2025,6,4]],"date-time":"2025-06-04T08:51:04Z","timestamp":1749027064000},"page":"841","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Intraoperative Confocal Laser Endomicroscopy Detects Prostate Cancer at the Single-Cell Level with High Specificity and in Real Time: A Preclinical Proof of Concept"],"prefix":"10.3390","volume":"18","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-5858-469X","authenticated-orcid":false,"given":"Ann-Christin","family":"Eder","sequence":"first","affiliation":[{"name":"Department of Nuclear Medicine, University Medical Center Freiburg, Faculty of Medicine, University of Freiburg, 79106 Freiburg, Germany"},{"name":"Division of Radiopharmaceutical Development, German Cancer Consortium (DKTK), Partner Site Freiburg, Freiburg, Germany and German Cancer Research Center, 69120 Heidelberg, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6560-2174","authenticated-orcid":false,"given":"Jessica","family":"Matthias","sequence":"additional","affiliation":[{"name":"Department of Optical Nanoscopy, Max Planck Institute for Medical Research, 69120 Heidelberg, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Francois","family":"Lacombe","sequence":"additional","affiliation":[{"name":"Mauna Kea Technologies, 75010 Paris, France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lisa-Charlotte","family":"Domogalla","sequence":"additional","affiliation":[{"name":"Department of Nuclear Medicine, University Medical Center Freiburg, Faculty of Medicine, University of Freiburg, 79106 Freiburg, Germany"},{"name":"Division of Radiopharmaceutical Development, German Cancer Consortium (DKTK), Partner Site Freiburg, Freiburg, Germany and German Cancer Research Center, 69120 Heidelberg, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Antoine","family":"Jacques","sequence":"additional","affiliation":[{"name":"Mauna Kea Technologies, 75010 Paris, France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nils","family":"Steinacker","sequence":"additional","affiliation":[{"name":"Department of Nuclear Medicine, University Medical Center Freiburg, Faculty of Medicine, University of Freiburg, 79106 Freiburg, Germany"},{"name":"Division of Radiopharmaceutical Development, German Cancer Consortium (DKTK), Partner Site Freiburg, Freiburg, Germany and German Cancer Research Center, 69120 Heidelberg, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Gaetan","family":"Christien","sequence":"additional","affiliation":[{"name":"Mauna Kea Technologies, 75010 Paris, France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Elodie","family":"Martin","sequence":"additional","affiliation":[{"name":"Mauna Kea Technologies, 75010 Paris, France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Aline","family":"Criton","sequence":"additional","affiliation":[{"name":"Mauna Kea Technologies, 75010 Paris, France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Matthias","family":"Eder","sequence":"additional","affiliation":[{"name":"Department of Nuclear Medicine, University Medical Center Freiburg, Faculty of Medicine, University of Freiburg, 79106 Freiburg, Germany"},{"name":"Division of Radiopharmaceutical Development, German Cancer Consortium (DKTK), Partner Site Freiburg, Freiburg, Germany and German Cancer Research Center, 69120 Heidelberg, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,6,4]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1545","DOI":"10.2967\/jnumed.117.191031","article-title":"PSMA Ligands for PET Imaging of Prostate Cancer","volume":"58","author":"Schwarzenboeck","year":"2017","journal-title":"J. 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